重组人内抑素对佐剂性关节炎大鼠成纤维样滑膜细胞内钙离子稳态的影响

目的 观察重组人内抑素(rhEndestatin)对佐剂性关节炎大鼠成纤维样滑膜细胞(AA FLSs)内钙离子(Ca2+)稳态的影响,探讨rhEndomafin促进AA FLSs凋亡的离子机制,为RA药物治疗及寻找其治疗新靶点提供实验依据.方法雄性SD大鼠12只,体质量140～160 g,分为正常组(n=3)和从模型组(n=9).模型组大鼠制备从模型,体外培养AA FLSs,应用Ca2+荧光指示剂Fluo-3/AM孵育培养的细胞,激光扫描共焦显微镜检测有、无细胞外Ca2+,而rhEndestatin作用所致AA FLSs胞内Ca2+荧光强度发生动态变化,可以判断rhEndostatin对AA FLSs胞内Ca2+浓度(Ca2+]I)的影响.结果在胞外有Ca2+的情况下,rhEndostatin可引起静态AA FLSCa2+];快速增加,rhEndostatin作用10s后,Ca2+];急剧增加达峰值,继之随时间缓慢下降,停止加药50 s后,Ca2+]I尚未回复到加药前基础水平;而在无细胞外钙环境中,rhEndostatin未引起AA FLSsCa2+]I变化.结论 rhEndestafin可促进AA FLSs胞外Ca2+内流,引起胞内Ca2+超载,从而促进从FLSs凋亡.

Effects of recombinant human endostatin on intracellular Ca2+ homeostasis in fibroblast-like synoviocytes in rats with adjuvant arthritis

Objective To observe the effects of recombinant human endostatin(rhEndostatin) on intracellular CaSUP>2+/SUP> homeostasis in fibroblast like synoviocytes(FLSs) in rats with adjuvant arthritis(AA), approach ionic mechanisms of AA FLSs apoptosis induced by rhEndostatin, and provide experimental data for possible drug treatment and their targets in rheumatoid arthritis (RA). Methods Twelve male Sprague Dawley(SD) rats weighing 140 160 g were used for these studies. The rats were divided at random into a normal group (EM>n/EM>=3) and an AA model group (EM>n/EM>=9). Adjuvant arthritis was induced in rats in the AA model group and the FLSs obtained from AA rats were cultured EM>in vitro/EM>. The cultured AA FLSs were incubated with the calcium ion sensitive fluorescent indicator fluo 3/AM. A confocal laser scanning microscope was used to measure changes of fluorescence intensity of intracellular CaSUP>2+/SUP> in the cells caused by rhEndostatin when the cells were immersed in buffer (with or without CaSUP>2+/SUP>), and to investigate the effect of rhEndostatin on intracellular free calcium concentration {［CaSUP>2+/SUP>］i} in AA FLS. Results Recombinant human endostatin could cause rapid ［CaSUP>2+/SUP>i elevation in static AA FLSs in CaSUP>2+/SUP>containing buffer. Within 10s after treatment with rhEndostatin, the elevated levels of ［CaSUP>2+/SUP>］i ran rapidly to peaked and was followed by a plateau period. The elevated ［CaSUP>2+/S